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AB273749

Human PDGFRB knockout SH-SY5Y cell line

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PDGFRB KO cell line available to order. KO validated by Western blot. Free of charge wild type control available. Knockout achieved by using CRISPR/Cas9, Homozygous: 5 bp deletion in exon 3. To order both knockout and wild-type control cells: select 2 x 1000000Cells/vial. To order only knockout cells: select 1000000Cells/vial.

View Alternative Names

Beta platelet derived growth factor receptor, Beta-type platelet-derived growth factor receptor, CD 140B, CD140 antigen-like family member B, CD140b antigen, IBGC4, IMF1, JTK12, OTTHUMP00000160528, PDGF Receptor beta, PDGF-R-beta, PDGFR, PDGFR 1, PDGFRB, PGFRB_HUMAN, Platelet derived growth factor receptor 1, Platelet derived growth factor receptor beta, Platelet derived growth factor receptor beta polypeptide

3 Images
Western blot - Human PDGFRB knockout SH-SY5Y cell line (AB273749)
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Western blot - Human PDGFRB knockout SH-SY5Y cell line (AB273749)

Lanes 1 - 4 : Merged signal (red and green). Green - ab69506 observed at 170 kDa. Red - loading control ab52866 (Rabbit anti-alpha Tubulin antibody [EP1332Y]) observed at 55 kDa.

ab69506 was shown to react with PDGFR beta in wild-type SH-SY5Y cells in Western blot with loss of signal observed in PDGFRB knockout cell line ab273749 (knockout cell lysate ab275523). Wild-type SH-SY5Y and PDGFRB knockout cell lysates were subjected to SDS-PAGE. Membranes were blocked in 3 % milk in TBS-T (0.1 % Tween®) before incubation with ab69506 and ab52866 (Rabbit anti-alpha Tubulin antibody [EP1332Y]) overnight at 4 ° at a 1 in 1000 dilution and a 1 in 20000 dilution respectively. Blots were incubated with Goat anti-Mouse IgG H&L (IRDye® 800CW) preabsorbed (ab216772) and Goat anti-Rabbit IgG H&L (IRDye® 680RD) preabsorbed (ab216777) secondary antibodies at 1 in 20000 dilution for 1 h at room temperature before imaging.

All lanes:

Western blot - Anti-PDGFR beta antibody [42G12] (<a href='/en-us/products/primary-antibodies/pdgfr-beta-antibody-42g12-ab69506'>ab69506</a>) at 1/1000 dilution

Lane 1:

Wild-type SH-SY5Y cell lysate at 30 µg

Lane 2:

PDGFRB knockout SH-SY5Y cell lysate at 30 µg

Lane 2:

Western blot - Human PDGFRB knockout SH-SY5Y cell line (ab273749)

Lane 3:

Human Skeletal Muscle tissue lysate at 30 µg

Lane 4:

HeLa cell lysate at 30 µg

Predicted band size: 123 kDa

Observed band size: 170 kDa

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Western blot - Human PDGFRB knockout SH-SY5Y cell line (AB273749)
  • WB

Lab

Western blot - Human PDGFRB knockout SH-SY5Y cell line (AB273749)

Lanes 1 - 4 : Merged signal (red and green). Green - ab32570 observed at 170 kDa. Red - loading control ab7291 (Mouse anti-Alpha Tubulin [DM1A]) observed at 55 kDa.

ab32570 was shown to react with PDGFRB in wild-type SH-SY5Y cells in Western blot with weakened signal observed in PDGFRB knockout cell line ab273749 (knockout cell lysate ab275523). Wild-type SH-SY5Y and PDGFRB knockout cell lysates were subjected to SDS-PAGE. Membranes were blocked in fluorescent western blot (TBS-based) blocking solution before incubation with ab32570 and ab7291 (Mouse anti-Alpha Tubulin [DM1A]) overnight at 4 ° at a 1 in 5000 dilution and a 1 in 20000 dilution respectively. Blots were incubated with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preabsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed (ab216776) secondary antibodies at 1 in 20000 dilution for 1 h at room temperature before imaging. The weak signal observed in PDGFRB knockout cell line should be PDGFRA.

All lanes:

Western blot - Anti-PDGFR alpha + PDGFR beta antibody [Y92] - C-terminal (<a href='/en-us/products/primary-antibodies/pdgfr-alpha-pdgfr-beta-antibody-y92-c-terminal-ab32570'>ab32570</a>) at 1/5000 dilution

Lane 1:

Wild-type SH-SY5Y cell lysate at 30 µg

Lane 2:

PDGFR beta knockout SH-SY5Y cell lysate at 30 µg

Lane 2:

Western blot - Human PDGFRB knockout SH-SY5Y cell line (ab273749)

Lane 3:

Human Skeletal Muscle tissue lysate at 30 µg

Lane 4:

HeLa cell lysate at 30 µg

Predicted band size: 123 kDa

Observed band size: 170 kDa

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Sanger Sequencing - Human PDGFRB knockout SH-SY5Y cell line (AB273749)
  • Sanger seq

Lab

Sanger Sequencing - Human PDGFRB knockout SH-SY5Y cell line (AB273749)

Allele-1 : 5 bp deletion in exon 3

Key facts

Cell type

SH-SY5Y

Species or organism

Human

Tissue

Bone marrow

Form

Liquid

form

Knockout validation

Sanger Sequencing,Western blot

Mutation description

Knockout achieved by using CRISPR/Cas9, Homozygous: 5 bp deletion in exon 3

Disease

Neuroblastoma

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Reactivity data

{ "title": "Reactivity Data", "filters": { "stats": ["", "Reactivity", "Dilution Info", "Notes"] }, "values": { "WB": { "reactivity":"TESTED_AND_REACTS", "dilution-info":"", "notes":"<p></p>" } } }
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Product details

We will provide viable cells that proliferate on revival.

This product is subject to limited use licenses from The Broad Institute and ERS Genomics Limited, and is developed with patented technology. For full details of the limited use licenses and relevant patents please refer to our limited use license and patent pages.

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What's included?

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Properties and storage information

Gene name
PDGFRB
Gene editing type
Knockout
Gene editing method
CRISPR technology
Knockout validation
Sanger Sequencing, Western blot
Zygosity
Homozygous
Shipped at conditions
Dry Ice
Appropriate short-term storage conditions
-196°C
Appropriate long-term storage conditions
-196°C
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Handling procedures

Initial handling guidelines

Upon arrival, the vial should be stored in liquid nitrogen vapor phase and not at -80°C. Storage at -80°C may result in loss of viability.

1. Thaw the vial in 37°C water bath for approximately 1-2 minutes.
2. Transfer the cell suspension (0.8 mL) to a 15 mL/50 mL conical sterile polypropylene centrifuge tube containing 8.4 mL pre-warmed culture medium, wash vial with an additional 0.8 mL culture medium (total volume 10 mL) to collect remaining cells, and centrifuge at 201 x g (rcf) for 5 minutes at room temperature. 10 mL represents minimum recommended dilution. 20 mL represents maximum recommended dilution.
3. Resuspend the cell pellet in 5 mL pre-warmed culture medium and count using a haemocytometer or alternative cell counting method seed all remaining cells into a T25.
4. Incubate the culture at 37°C incubator with 5% CO2. Check the culture one day after revival and continue to check until 80% confluent. Media change can be given if needed.
5. Once confluent passage into an appropriate flask at a density of 2x104 cells/cm2. Seeding density is given as a guide only and should be scaled to align with individual lab schedules. Cultures should be monitored daily.

Subculture guidelines
  • All seeding densities should be based on cell counts gained by established methods.
  • These cells grow as a mixture of floating and adherent cells.
  • Remove media containing floating cells and recover cells by centrifugation, detach cells using standard methods, combine with floating cells and transfer to a new culture flask.
  • A guide seeding density of 2x104 cells/cm2 is recommended.
  • Cells should be seeded at a density conducive to cell–cell communication to proliferate. If cells are seeded too sparsely, growth rate is reduced and cell death is high.
Culture medium

1:1 mixture of EMEM and F-12K + 10% FBS

Cryopreservation medium

Cell Freezing Medium-DMSO Serum free media, contains 8.7% DMSO in MEM supplemented with methyl cellulose.

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Supplementary information

This supplementary information is collated from multiple sources and compiled automatically.

PDGFR beta also known as PDGFR-b or PDGFRB is a cell surface tyrosine kinase receptor with a molecular mass of about 180 kDa. It binds the PDGF (platelet-derived growth factor) family of ligands. PDGFR beta is commonly found in various tissues including blood vessels and connective tissues and is highly expressed in cells like pericytes and fibroblasts. The receptor plays a critical role in cell signaling mechanisms involving proliferation chemotaxis and survival.
Biological function summary

The receptor plays an essential role in the regulation of cell growth and development. PDGFR beta undergoes dimerization and autophosphorylation upon ligand binding initiating a series of downstream signaling cascades. This receptor is often part of a complex with other receptor proteins promoting interactions necessary for signal propagation. Its main biological functions include mediating cellular responses to environmental signals that contribute to tissue repair and angiogenesis.

Pathways

PDGFR beta is an important player within the PI3K-Akt and MAPK signaling pathways. It works alongside proteins such as PI3K and Ras to regulate cellular responses related to growth and survival. These pathways facilitate cross-talk with other cellular processes influencing various cellular outcomes. This receptor's activity regulates critical physiological functions by providing signals that maintain cellular homeostasis under various physiological conditions.

PDGFR beta has significant implications in the progression of cancer and fibrotic diseases. Its overexpression or mutation can lead to anomalous signaling that contributes to tumorigenesis particularly in connective tissue tumors known as sarcomas. Additionally PDGFR beta's role in promoting fibroblast activity makes it relevant in fibrotic diseases such as pulmonary fibrosis. Abnormal activation of PDGFR beta can interact with related proteins like VEGF receptors enhancing pathogenic responses and contributing to disease severity.
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Quality control

STR analysis

CSF1PO, D13S317, D7S820, D5S818, TH01, D16S539, TPOX

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Cell culture

Biosafety level

EU: 1 US: 1

Adherent/suspension

Adherent

Gender

Female

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Product protocols

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Complementary Products

Primary Antibodies

AB32570

Anti-PDGFR alpha + PDGFR beta antibody [Y92] - C-terminal

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Western blot - Anti-PDGFR alpha + PDGFR beta antibody [Y92] - C-terminal (AB32570)

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Primary Antibodies

AB203491

Anti-PDGFR alpha antibody [EPR22059-270]

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PDGFR alpha antibody [EPR22059-270] (AB203491)

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Cell Lines & Lysates

AB280041

Human SNAP25 knockout SH-SY5Y cell line

Advanced Validation

Western blot - Human SNAP25 knockout SH-SY5Y cell line (AB280041)

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Secondary Antibodies

AB150113

Goat Anti-Mouse IgG H&L (Alexa Fluor® 488)

Immunocytochemistry/ Immunofluorescence - Goat Anti-Mouse IgG H&L (Alexa Fluor® 488) (AB150113)

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Product promise

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